Funded Research Projects

Zhou Lab, University of Pennsylvania

“Modeling CDKL5 Related Disorders in Mice”

Researchers: Zhaolan Zhou, PhD, started 2014: The development of mice with CDKL5 is essential to clinically study the effects that various drugs would have in order to move research into clinical trials. Dr. Zhou’s lab has created several different types of mice with CDKL5 mutations which allows us to study the molecular mechanisms that are important in the disorder and to evaluate how different types of mutations cause different types of effects. These models have started to identify specific molecular pathways involved in CDKL5. This project includes the creation of a reversible mouse model. Utilizing these models, Dr. Zhou has started several trials with compounds that may help alleviate some of symptoms of CDKL5.

“CDKL5 Mouse Model (Knock-In)”

Researchers: Zhaolan Zhou, PhD, started 2011: Project: IFCR has funded a knock-in mouse model with Dr. Zhaolan (Joe) Zhou at the University of Pennsylvania. He is developing a germline transmitted knock-in mouse which will have a nonsense mutation R59X. It is imperative that different types of mutated mice are developed to be able to further develop studies to determine what the CDKL5 protein really does and to see how new drugs affect different types of mutations. This particular nonsense mutation was selected for a few reasons. It appears to be a more common mutation for CDKL5 with 5 reported cases in the literature, it is at a strategic site in the gene and we have correlative samples from both a male and female affected with CDKL5 in which iPS cells are also being developed.

Biotech companies are working to develop read-through compounds which will allow the production of RNA and protein despite having a nonsense mutation. Many of the CDKL5 affected children have this type of mutation. These studies will help to correlate activities in mouse and human cells and will help identify a reproducible cellular phenotype that would allow us to test the read-through compounds that are already available. With these correlations, it will be much easier to take these compounds into clinical trials.

In Progress: Continued breeding

Phenotype characterization (Anticipate 2013)

Attempt to reverse the phenotype (symptoms of CDKL5) in mice with currently available “read-through” drugs.

Need 2-3 other mice created with different “human” mutations to allow better phenotype characterization and allow for best model prediction of drug discovery and modifier genes

Correlation of mouse neuronal development with iPS neuronal development

“Development of Antibodies Against CDKL5”

Researchers: Zhaolan Zhou, PhD, started 2012

Muotri Lab, University of California San Diego

“Modeling CDKL5 Syndrome Using iPSC-Derived Human Neurons”

Researcher: Alysson Muotri, PhD, started 2014: Utilizing advanced technology this lab has successfully been able to grow neurons directly from several children with CDKL5 from s simple skin biopsy. These CDKL5 neurons allows for many areas of investigation such as looking at pathways involved in CDKL5 disorder, including some of the pathways that may cause seizures. Using single cell gene expression profiling it has been discovered that CDKL5 neurons express a unique set of neurotransmitters. It has also been discovered that there is a unique change that happens after cells are stimulated. This information allows this technology to be used to identify how different drugs affect the function and growth of the neurons, and this will lead to the discovery of various medications that can help change the outcome of CDKL5. Also, the cells will be specially screened to evaluate a specific molecular profile of CDKL5 cells, which in turn will help to determine other pathways involved with CDKL5 and could lead to more therapeutic targets.

“Modeling CDKL5 Syndrome using human Neurons”

Researchers: Alysson Muotri, Ph.D and Priscilla Negraes, Ph.D, started September 2012: IFCR has funded an 2-year postdoctoral position for “modeling CDKL5 syndrome using human neurons”. The goal is to examine specific neuronal gene expression profiles, with the aim to accelerate the discovery of novel therapeutic drugs to treat CDKL5.

“Development of iPS cells and Differentiation into Neurons”

Researcher: Alysson Muotri, PhD Started: April 2011: Create induced pluripotent stem cells from skin fibroblasts of patients with CDKL5 and differentiate into neuronal cells. Update (May 2013): Multiple iPS cell lines have been successfully created and differentiated into CDKL5 neurons from males with CDKL5, and their corresponding controls. In addition, we have one cell line from a female with a nonsense mutation R59X, and one from a male with the same mutation. In Progress:

· Characterization of cell lines

· Correlation to mouse lines

· Drug screening and molecular profiling Future Studies

Required: May need more females Need to develop lines with variety of mutations

Benke Lab, University of Colorado Denver

“Investigation of the role of CDKL5 expression, localization and function in seizures and intellectual disability”

Researchers: Heather Caballes, PhD and Tim Benke, MD, PhD, started 2015: CDKL5 plays roles in plasticity and development, and its disruption leads to syndromes characterized by seizures and intellectual disability; however, the exact mechanisms that lead to the development of epilepsy and intellectual disability are unknown. The experiments proposed here will begin to elucidate these mechanisms by characterizing expression of CDKL5 in human epileptic tissue and in a rat model of early life seizures, as well as examine CDKL5 role in AMPA receptor desensitization using CDKL5 KO mice

“Children’s Hospital Colorado CDKL5 Center of Excellence”

Researcher: Tim Benke, MD, Ph.D , started April 2013: CDKL5 Centers for Excellence are the only multidisciplinary clinics for the treatment of people, a majority of whom are children, with a CDKL5 Disorder (CDKL5), also serving as clinical research consortiums.

In progress

European Molecular Biology Laboratory, Monterotondo, Italy

“CDKL5 Mouse Model (Knock out)”

Researchers: Cornelius Gross, PhD and Elena Amendola, PhD- post doctoral fellow, started 2010 and completed 2012: Create CDKL5 Knockout mouse model (Generate a mouse that lacks the Cdkl5 gene). Other labs have said that CDKL5 KO mice were embryonic lethal so this lab is using a specialized Cr:e-lox technique to create mice derived entirely from the knockout ES cells, while in the traditional procedure only part of the offspring are derived form the knockout ES cells.

Telethon Kids Institute, Perth, Australia

“CDKL5 Database Registry” Ongoing support

Researchers: Dr. Helen Leonard and Dr. Jenny Downs, continued globally from original 2012 grant: An extremely important aspect of CDKL5 research is to gain an understanding of the effects and changes that happen over time- often referred to as the “Natural History” of the disorder. This group has been studying the epidemiology of neurological disorders for many years, including Rett Syndrome and Down Syndrome. Utilizing a questionnaire developed specifically for CDKL5 we are learning so much about the spectrum of the disorder and identifying patterns of treatments and interventions that may lead to an improved quality of life for our children. Dr. Leonard and Dr. Jenny Downs work in collaboration with the CDKL5 centers of Excellence and many families and clinicians around the world. Please visit https://www.cdkl5.com/Research/Database.aspx to participate.

Researchers: Stephanie Fehr and Dr. Helen Leonard, started 2012: As part of the Natural History of CDKL5 disorder, the Australian pediatric surveillance study will help determine the incidence and prevalence CDKL5 in Australia, thereby highlighting the impact CDKL5 disorder has on families, caregivers and society.

Cleveland Clinic

“Cleveland Clinic CDKL5 Center of Excellence”

Researcher: Dr. Sumit Parikh, MD, started 2014: CDKL5 Centers for Excellence are the only multidisciplinary clinics for the treatment of people, a majority of whom are children, with a CDKL5 Disorder (CDKL5), also serving as clinical research consortiums.

In progress.

Boston Children’s Hospital

“Boston Children’s Hospital Center of Excellence”

Researcher: Heather Olson, MD, started 2013: CDKL5 Centers for Excellence are the only multidisciplinary clinics for the treatment of people, a majority of whom are children, with a CDKL5 Disorder (CDKL5), also serving as clinical research consortiums.

In progress.

University of Bologna, Italy

“Drug Therapy Targeted to Core Molecules in Neural Plasticity Cascades: A Promising Tool for the CDKL5 Variant of Rett Syndrome” (Co-Funded with Rettsyndrome.org)

Researchers: Claudia Fuchs, PhD., started 2014: CDKL5 plays a role in a neurologic function called “plasticity” which is important for the multiple aspects of learning and memory. There are many different mechanisms that play a role in plasticity and this lab has identified some of the key pathways that involve CDKL5. Current studies include exploring existing drugs which may allow restoration of plasticity in CDKL5. If these studies show promise this would allow for a fairly rapid transition to a clinical trial.

Institute of Neuroscience, National Cheng-Chi University, Taiwan

“Neurochemical and Therapeutic Studies in Mouse Models of CDKL5 Disorder”

Researchers: Wenlin Liao, PhD, started 2015: Mice with CDKL5 show impaired social communication and enhanced stereotypy which are key features of autism. These mice also develop increased impulsivity and locomotor activity, simulating features of Attention Deficit Hyperactivity Disorder (ADHD). This lab is examining how CDKL5 is causing these disorders and they have some very interesting results showing new pathways in which CDKL5 is involved. Utilizing these discoveries, they will be studying therapies that may help to alleviate these problems.

Tao Lab, Purdue University

“Identification of CDKL5 direct substrates based on kinase linked phosphoproteomics”

Researchers: Weiguo Andy Tao, PhD, started 2015: CDKL5 is a serine/threonine kinase but the complete characterization of its targets are lacking. Further identification of protein targets for this kinase is necessary to provide more detailed biologic effects of CDKL5. This project is using kinase linked phosphoproteomics to further identify the phosphorylation targets of CDKL5.

University of Turin and National Institute of Neuroscience

Researchers: Maurizio Giustetto, PhD and Tommaso Pizzorusso, PhD, started 2015: This project will further explore CDKL5 roles in synaptic function and plasticity concentrating on therapeutic potentials with agents that can enhance this pathway and specifically drugs to increase activity of the AKT/mTOR pathway. This project will utilize use of the mouse models and will also include the evaluation of new antibody formation.

University of Insubria, Italy

“Investigation of the importance of an uncharacterized MeCP2 phospho-isoform for neuronal morphogenesis and chromatin related functions, and the relationship between CDKL5 and MeCP2”

Researchers: Charlotte Kilstrup-Nielsen, PhD, started 2013:

Pyschogenics, Inc., New York

“Phenotyping of a CDKL5 Mouse Model”

The goal of the study is to assess the phenotype of a mouse models of CDKL5 with the longer-term goal of setting up a drug screen battery.

University of California San Francisco; National Institute for Medical Research, London UK

“Identification of CDKL5’s Phosphorylation Targets”

Researchers: Sila Konur Ultanir, Ph.D, started 2012: Dr. Ultanir is using a recently developed kinase substrate identification method to reveal the phosphorylation sites of CDKL5. The goal is to gain valuable information and a better understanding of the synaptic and molecular functions of CDKL5.